LOCUS       V039078                 8861 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V039078
VERSION     V039078
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             935..1651
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK"
                     /label="EGFP"
     CDS             1658..4351
                     /label="ILF3(NM_017620)"
                     /note="ILF3(NM_017620)"
                     /gene="ILF3"
     CDS             4358..4423
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    4461..4685
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4731..5159
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5173..5502
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5569..6363
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    6537..6658
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6707..6723)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6731..6747
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(6755..6785)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6800..6821
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7109..7694)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7865..8725)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(8726..8830)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccatggtg agcaagggcg aggagctgtt
      961 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     1021 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
     1081 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt
     1141 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     1201 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     1261 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     1321 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
     1381 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg
     1441 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     1501 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag
     1561 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     1621 gatcactctc ggcatggacg agctgtacaa ggaattcatg cgtccaatgc gaatttttgt
     1681 gaatgatgac cgccatgtga tggcaaagca ttcttccgtt tatccaacac aagaggagct
     1741 ggaggcagtc cagaacatgg tgtcccacac ggagcgggcg ctcaaagctg tgtccgactg
     1801 gatagacgag caggaaaagg gtagcagcga gcaggcagag tccgataaca tggatgtgcc
     1861 cccagaggac gacagtaaag aaggggctgg ggaacagaag acggagcaca tgaccagaac
     1921 cctgcgggga gtgatgcggg tgggcctggt ggcaaagggc ctcctactca agggggactt
     1981 ggatctggag ctggtgctgc tgtgtaagga gaagcccaca accgccctcc tggacaaggt
     2041 ggccgacaac ctggccatcc agcttgctgc tgtaacagaa gacaagtacg aaatactgca
     2101 atctgtcgac gatgctgcga ttgtgataaa aaacacaaaa gagcctccat tgtccctgac
     2161 catccacctg acatcccctg ttgtcagaga agaaatggag aaagtattag ctggagaaac
     2221 gctatcagtc aacgaccccc cggacgttct ggacaggcag aaatgccttg ctgccttggc
     2281 gtccctccga cacgccaagt ggttccaggc cagagccaac gggctgaagt cttgtgtcat
     2341 tgtgatccgg gtcttgaggg acctgtgcac tcgcgtgccc acctggggtc ccctccgagg
     2401 ctggcctctc gagctcctgt gtgagaaatc cattggcacg gccaacagac cgatgggtgc
     2461 tggcgaggcc ctgcggagag tgctggagtg cctggcgtcg ggcatcgtga tgccagatgg
     2521 ttctggcatt tatgaccctt gtgaaaaaga agccactgat gctattgggc atctagacag
     2581 acagcaacgg gaagatatca cacagagtgc gcagcacgca ctgcggctcg ctgccttcgg
     2641 ccagctccat aaagtcctag gcatggaccc tctgccttcc aagatgccca agaaaccaaa
     2701 gaatgaaaac ccagtggact acaccgttca gatcccacca agcaccacct atgccattac
     2761 gcccatgaaa cgcccaatgg aggaggacgg ggaggagaag tcgcccagca aaaagaagaa
     2821 gaagattcag aagaaagagg agaaggcaga gcccccccag gctatgaatg ccctgatgcg
     2881 gttgaaccag ctgaagccag ggctgcagta caagctggtg tcccagactg ggcccgtcca
     2941 tgcccccatc tttaccatgt ctgtggaggt tgatggcaat tcattcgagg cctctgggcc
     3001 ctccaaaaag acggccaagc tgcacgtggc cgttaaggtg ttacaggaca tgggcttgcc
     3061 gacgggtgct gaaggcaggg actcgagcaa gggggaggac tcggctgagg agaccgaggc
     3121 gaagccagca gtggtggccc ctgccccagt ggtagaagct gtctccaccc ctagtgcggc
     3181 ctttccctca gatgccactg ccgagaacgt aaaacagcag gggccgatcc tgacaaagca
     3241 cggcaagaac ccagtcatgg agctgaacga gaagaggcgt gggctcaagt acgagctcat
     3301 ctccgagacc gggggcagcc acgacaagcg cttcgtcatg gaggtcgaag tggatggaca
     3361 gaagttccaa ggtgctggtt ccaacaaaaa ggtggcgaag gcctacgctg ctcttgctgc
     3421 cctagaaaag cttttccctg acacccctct cgcccttgat gccaacaaaa agaagagagc
     3481 cccagtaccc gtcagagggg gaccgaaatt tgctgctaag ccacataacc ctggcttcgg
     3541 catgggaggc cccatgcaca acgaagtgcc cccacccccc aaccttcgag ggcggggaag
     3601 aggcgggagc atccggggac gagggcgcgg gcgaggattt ggtggcgcca accatggagg
     3661 ctacatgaat gccggtgctg ggtatggaag ctatgggtac ggaggcaact cggcgacagc
     3721 aggctacagt cagttctaca gcaacggagg gcattctggg aatgccagtg gcggtggcgg
     3781 cgggggcggt ggtggctcct ccggctatgg ctcctactac caaggtgaca actacaactc
     3841 accggtgccc ccaaaacacg ctgggaagaa gcagccgcac gggggccagc agaagccctc
     3901 ctacggctcg ggctaccagt cccaccaggg ccagcagcag tcctacaacc agagccccta
     3961 cagcaactat ggccctccac agggcaagca gaaaggctat aaccatggac aaggcagcta
     4021 ctcctactcg aactcctaca actctcccgg gggcgggggc ggatccgact acaactacga
     4081 gagcaaattc aactacagtg gtagtggagg ccgaagcggc gggaacagct acggctcagg
     4141 cggggcatcc tacaacccag ggtcacacgg gggctacggc ggaggttctg ggggcggctc
     4201 ctcataccaa ggcaaacaag gaggctactc acagtcgaac tacaactccc cggggtccgg
     4261 ccagaactac agtggccctc ccagctccta ccagtcctca caaggcggct atggcagaaa
     4321 cgcagaccac agcatgaact accagtacag actcgaggac tacaaagacc atgacggtga
     4381 ttataaagat catgacatcg actacaagga tgacgatgac aagtagtgag ggcccgttta
     4441 aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     4501 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     4561 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     4621 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     4681 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
     4741 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     4801 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     4861 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     4921 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     4981 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     5041 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     5101 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     5161 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     5221 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     5281 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     5341 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     5401 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     5461 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     5521 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     5581 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     5641 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     5701 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     5761 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     5821 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     5881 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     5941 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     6001 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     6061 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     6121 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     6181 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     6241 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     6301 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     6361 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     6421 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     6481 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     6541 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     6601 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     6661 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     6721 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     6781 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     6841 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     6901 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     6961 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     7021 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     7081 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     7141 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     7201 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     7261 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     7321 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     7381 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     7441 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     7501 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     7561 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     7621 gcaaacaaac caccgctggt agcggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     7681 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     7741 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     7801 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     7861 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     7921 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     7981 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     8041 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     8101 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     8161 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     8221 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     8281 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     8341 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     8401 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     8461 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     8521 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     8581 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     8641 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     8701 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     8761 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     8821 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt c
//
